We propose a comprehensive program to determine whether human cancers contain sequences representing 5 distinct human papillomaviruses (HPV-1 to HPV-5). The possible role of HPVs in human cancer is an open question and recent findings suggest that the carcinogenic properties of papillomaviruses have been underestimated. Several animal cancers are known to be caused by papillomaviruses, and several types of human papillomatous lesions have been reported to progress to malignancy. We also propose to study several aspects of the molecular biology and life cycle of these viruses that are relevant to further understanding their oncogenic properties. Our specific aims are: (1) to clone the DNAs of 5 distinctly different human papillomaviruses (HPV), HPV-1, HPV-2, HPV-3, HPV-4, and HPV-5, as well as other new isolates since HPVs have not been grown in culture and HPV DNAs are needed in quantity for cancer analyses and molecular studies; (2) to characterize the cloned HPV DNAs by cross hybridization and heteroduplex mapping under different stringency conditions, and by restriction map analysis; (3) to perform extensive human cancer studies (1069 cancers and 276 high molecular weight DNAs are available) with representative HPV probes under conditions that detect 0.01-0.1 copies of HPV specific DNA and RNA per cell; (4) to analyze benign papillomatous tumors for content of specific HPV DNA sequences and possible integration and expression of the viral genome since papillomatous tumors provide a unique and potentially powerful system to analyze virus-cell interaction and possibly virus-cell transformation under natural conditions, and possible specific sites of integration on cell DNA and viral DNA; and (5) to develop systems to study the organization, life cycle, and expression of putative transforming functions of HPV genomes by transformation-transfection of appropriate cells with cloned HPV DNA and restriction fragments. This research is an ongoing collaboration between Drs. Maurice Green and William Wold at the Institute for Molecular Virology (IMV) and Dr. Gerard Orth (and his collaborators Drs. Michel Favre and Odile Croissant) at the Pasteur Institute.